cAMP Signaling, Softcover reprint of the original 1st ed. 2015 Methods and Protocols Methods in Molecular Biology Series, Vol. 1294

This detailed volume encompasses new technological developments that specifically address questions related to adenosine 3?,5?-monophosphate (cAMP) compartmentalization, that probe relevant protein-protein interactions, that increase the spatial and temporal resolution of cAMP signal detection, and that can facilitate integration of the mounting complexity of the information that is becoming available on this signaling system. cAMP, the prototypical intracellular second messenger, regulates a large variety of cellular functions and biological processes, including gene transcription, cell metabolism, proliferation, development, as well as more specialized functions depending on the cell type, so the realization of its extremely complex spatial organization and local regulation is providing novel mechanistic insight into cell physiology and is producing a novel framework for the identification of disease mechanisms. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Practical and authoritative, cAMP Signaling: Methods and Protocols serves as a vital resource for researchers working in this expanding, complex field.

1. Simultaneous Assessment of cAMP Signaling Events in Different Cellular Compartments Using FRET-Based Reporters

            Alex Burdyga and Konstantinos Lefkimmiatis

2. Recording Intracellular cAMP Levels with EPAC-Based FRET Sensors by Fluorescence Lifetime Imaging

            Marcel Raspe, Jeffrey Klarenbeek, and Kees Jalink

3. A Novel Approach Combining Real-Time Imaging and the Patch-Clamp Technique to Calibrate FRET-Based Reporters for cAMP in Their Cellular Microenvironment

            Andreas Koschinski and Manuela Zaccolo

4. Structure-Based, In Silico Approaches for the Development of Novel cAMP FRET Reporters

            Matías Machado and Sergio Pantano

5. Automated Image Analysis of FRET Signals for Subcellular cAMP Quantification

            Silas J. Leavesley, Arie Nakhmani, Yi Gao, and Thomas C. Rich

6. Channel-Based Reporters for cAMP Detection

            Thomas C. Rich, Wenkuan Xin, Silas J. Leavesley, and Mark Taylor

7. Imaging Sub-Plasma Membrane cAMP Dynamics with Fluorescent Translocation Reporters

            Anders Tengholm and Olof Idevall-Hagren

8. Adenoviral Transduction of FRET-Based Biosensors for cAMP in Primary Adult Mouse Cardiomyocytes

            Oliver Lomas, Marcella Brescia, Ricardo Carnicer, Stefania Monterisi, Nicoletta Surdo, and Manuela Zaccolo

9. Generation of Transgenic Mice Expressing FRET Biosensors

            Daniela Hübscher and Viacheslav O. Nikolaev

10. Photoactivatable Adenylyl Cyclase (PACs) as a Tool to Study cAMP Signaling In Vivo: An Overview

            Marina Efetova and Martin Schwärzel

11. Selective Disruption of the AKAP Signaling Complexes

            Eileen J. Kennedy and John D. Scott

12. Screening for Small Molecule Disruptors of AKAP-PKA Interactions

            Carolin Schächterle, Frank Christian, João Miguel Parente Fernandes, Walter Rosenthal, and Enno Klussmann

13. Structure-Based Bacteriophage Screening for AKAP-Selective PKA Regulatory Subunit Variants

            Ryan Walker-Gray and Matthew G. Gold

14. A Yeast-Based High-Throughput Screen for Modulators of Phosphodiesterase Activity

            Ana Santos de Medeiros and Charles S. Hoffman

15. Separation of PKA and PKG Signaling Nodes by Chemical Proteomics

            Eleonora Corradini, Albert J.R. Heck, and Arjen Scholten

16. Development of Computational Models of cAMP Signaling

            Susana R. Neves-Zaph and Roy S. Song

Includes cutting-edge methods and protocols on cAMP signaling

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from experts